Investigation of fragmentation pathways of protonated 2-methoxypyrimidine derivatives

Several representative pyrimidine derivatives were selected to undergo electrospray ionization (ESI) followed by collision-induced dissociation tandem mass spectrometry (CID MS/MS) experiments. Two competitive pathways were found to govern the formation of major fragment ions from protonated species of these molecules. The pathways were largely affected by the 2-O-methyl group but not significantly influenced by the substitution on C-5 site of the pyrimidine ring. These findings were supported by both deuterium labeling CID MS/MS experiments and theoretical calculations. The deuterium labeled pyrimidine ion molecules were generated in-source in ESI from the fully deuterated hydrazinyl pyrimidines, which were readily obtained through hydrogen/deuterium (H/D) exchange when dissolved in deuterium oxide (D₂O).